Hydrophilic light absorbing compositions for determination of physiological function in critically ill patients

ABSTRACT

Highly hydrophilic indole and benzoindole derivatives that absorb and fluoresce in the visible region of light are disclosed. These compounds are useful for physiological and organ function monitoring. Particularly, the molecules of the invention are useful for optical diagnosis of renal and cardiac diseases and for estimation of blood volume in vivo.

RELATED APPLICATIONS

This application is the first of two Continuations, each filed Jul. 16,2007, and each a Continuation of pending U.S. patent application Ser.No. 10/744,334, filed on Dec. 23, 2003, which is a Continuation-In-Partof U.S. patent application Ser. No. 09/688,943, filed on Oct. 16, 2000,now U.S. Pat. No. 6,669,926, the disclosures of each hereby incorporatedby reference in its entirety.

TECHNICAL FIELD

Novel optical probes for use in physiological function monitoring,particularly indole and benzoindole compounds.

BACKGROUND OF THE INVENTION

Dynamic monitoring of physiological functions of patients at the bedsideis highly desirable in order to minimize the risk of acute renal failurebrought about by various clinical, physiological, and pathologicalconditions (Rabito et al., Renal function in patients at risk withcontrast material-induced acute renal failure: Noninvasive real-timemonitoring, Radiology 1993, 186, 851; Tilney and Lazarus, Acute renalfailure in surgical patients: Causes, clinical patterns, and care,Surgical Clinics of North America, 1983, 63, 357; VanZee et al., Renalinjury associated with intravenous pyelography in non-diabetic anddiabetic patients, Annals of Internal Medicine, 1978, 89, 51-54;Lundqvist et al., Iohexol clearance for renal function measurement ingynecologic cancer patients, Acta Radiologica, 1996, 37, 582; Guesry etal., Measurement of glomerular filtration rate by fluorescent excitationof non-radioactive meglumine iothalamate, Clinical Nephrology, 1975, 3,134). This monitoring is particularly important in the case ofcritically ill or injured patients because a large percentage of thesepatients face the risk of multiple organ failure (MOF), resulting indeath (Baker et al., Epidemiology of Trauma Deaths, American Journal ofSurgery, 1980, 144; Regel et al., Treatment Results of Patients withMultiple Trauma: An Analysis of 3406 Cases Treated Between 1972 and 1991at a German Level I Trauma Center, Journal of Trauma, 1995, 38, 70). MOFis a sequential failure of lung, liver, and kidneys, and is incited byone or more severe causes such as acute lung injury (ALI), adultrespiratory distress syndrome (ARDS), hypermetabolism, hypotension,persistent inflammatory focus, or sepsis syndrome. The commonhistological features of hypotension and shock leading to MOF includetissue necrosis, vascular congestion, interstitial and cellular edema,hemorrhage, and microthrombi. These changes affect the lung, liver,kidneys, intestine, adrenal glands, brain, and pancreas, in descendingorder of frequency (Coalson, Pathology of Sepsis, Septic Shock, andMultiple Organ Failure, New Horizons: Multiple Organ Failure Syndrome,Bihari and Cerra (Eds.), Society of Critical Care Medicine, FullertonCalif., 1986, pp. 27-59). The transition from early stages of trauma toclinical MOF is marked by the extent of liver and renal failure and achange in mortality risk from about 30% to about 50% (Cerra, MultipleOrgan Failure Syndrome. New Horizons: Multiple Organ Failure, Bihari andCerra (Eds). Society of Critical Care Medicine, Fullerton Calif., 1989,pp. 1-24).

Serum creatinine measured at frequent intervals by clinical laboratoriesis currently the most common way of assessing renal function andfollowing the dynamic changes in renal function which occur incritically ill patients (Doolan et al., A clinical appraisal of theplasma concentration and endogenous clearance of creatinine, AmericanJournal of Medicine, 1962, 32, 65-79; J. B. Henry (Ed). ClinicalDiagnosis and Management by Laboratory Methods, 17th Edition, W. B.Saunders, Philadelphia Pa., 1984); Speicher, The right test: Aphysician's guide to laboratory medicine, W. B. Saunders, PhiladelphiaPa., 1989). These values are frequently misleading, since age, state ofhydration, renal perfusion, muscle mass, dietary intake, and many otherclinical and anthropometric variables affect the value. In addition, asingle value returned several hours after sampling is difficult tocorrelate with other important physiologic events such as bloodpressure, cardiac output, state of hydration and other specific clinicalevents (e.g., hemorrhage, bacteremia, ventilator settings and others).An approximation of glomerular filtration rate can be made via a 24-hoururine collection, but this requires 24 hours to collect the sample,several more hours to analyze the sample, and a meticulous bedsidecollection technique. New or repeat data are equally cumbersome toobtain. Occasionally, changes in serum creatinine must be furtheradjusted based on the values for urinary electrolytes, osmolality, andderived calculations such as the “renal failure index” or the“fractional excretion of sodium.” These require additional samples ofserum collected contemporaneously with urine samples and, after a delay,precise calculations. Frequently, dosing of medication is adjusted forrenal function and thus can be equally as inaccurate, equally delayed,and as difficult to reassess as the values upon which they are based.Finally, clinical decisions in the critically ill population are oftenas important in their timing as they are in their accuracy.

Exogenous markers such as inulin, iohexol, ⁵¹Cr-EDTA, Gd-DTPA, or^(99m)Tc-DTPA have been reported to measure the glomerular filtrationrate (GFR) (Choyke et al., Hydrated clearance of gadolinium-DTPA as ameasurement of glomerular filtration rate, Kidney International, 1992,41, 1595; Tweedle et al., A noninvasive method for monitoring renalstatus at bedside, Invest. Radiol., 1997, 32, 802; Lewis et al.,Comparative evaluation of urograhic contrast media, inulin, and ^(99m)Tc-DTPA clearance methods for determination of glomerular filtrationrate in clinical transplantation, Transplantation, 1989, 48, 790). Othermarkers such as ¹²³I and ¹²⁵I labeled o-iodohippurate or ^(99m)Tc-MAG₃are used to assess tubular secretion process (Tauxe, Tubular Function,in Nuclear Medicine in Clinical Urology and Nephrology, Tauxe andDubovsky, Editors, p. 77, Appleton Century Crofts, East Norwalk, 1985;Muller-Suur, and Muller-Suur, Glomerular filtration and tubularsecretion of MAG ₃ in rat kidney, Journal of Nuclear Medicine, 1989, 30,1986). However, these markers have several undesirable properties suchas the use of radioactivity or ex-vivo handling of blood and urinesamples. Thus, in order to assess the status and to follow the progressof renal disease, there is a considerable interest in developing asimple, safe, accurate, and continuous method for determining renalfunction, preferably by non-radioactive procedures. Other organs andphysiological functions that would benefit from real-time monitoringinclude the heart, the liver, and blood perfusion, especially in organtransplant patients.

Hydrophilic, anionic substances are generally recognized to be excretedby the kidneys (Roch-Ramel et al., Renal excretion and tubular transportof organic anions and cations, Handbook of Physiology, Section 8,Neurological Physiology, Vol. II, Windhager, Ed., p. 2189, OxfordUniversity Press, New York, 1992; Nosco and Beaty-Nosco, Chemistry oftechnetium radiopharmaceuticals 1: Chemistry behind the development oftechnetium-^(99m) compounds to determine kidney function, CoordinationChemistry Reviews, 1999, 184, 91). It is further recognized that drugsbearing sulfonate residues exhibit improved clearance through thekidneys (Baldas and Bonnyman, Preparation, HPLC studies and biologicalbehavior of techentium-^(99m) and ^(99m) TcN0-radiopharmaceuticals basedon quinoline type ligands, Nucl. Med. Biol., 1999, 19, 491; Hansen etal., Synthesis of the sulfonate and phosphonate derivatives ofmercaptoacetyltriglycine. X-ray crystal structure of Na ₂[ReO(mercaptoacetylglycylglycylaminomethane-sulfonate)].3H ₂0,Met.-Based Drugs, 1994, 1, 31).

Assessment of renal function by continuously monitoring the bloodclearance of exogenous optical markers, viz., fluorescein bioconjugatesderived from anionic polypeptides, has been developed by us and byothers (Dorshow et al., Noninvasive fluorescence detection of hepaticand renal function, J. Biomedical Optics, 1998, 3, 340; Sohtell et al.,FITC-Inulin as a Kidney Tubule Marker in the Rat, Acta. Physiol. Scand.,1983, 119, 313, each of which is expressly incorporated herein byreference). The main drawback of high molecular weight polypeptides isthat they are immunogenic. In addition, large polymers with narrowmolecular weight distribution are difficult to prepare, especially inlarge quantities. Thus, there is a need in the art to develop lowmolecular weight compounds that absorb and/or emit light that can beused for assessing renal, hepatic, cardiac and other organ functions.

SUMMARY

The present invention overcomes these difficulties by incorporatinghydrophilic anionic or polyhydroxy residues in the form of sulfates,sulfonates, sulfamates, phosphates, polyethers, polyamino carboxylicacids and their derivatives, and strategically positioned hydroxylgroups. Thus, the present invention is related to novel dyes containingmultiple hydrophilic moieties and their use as diagnostic agents forassessing organ function and functional status of tumors.

The novel compounds of the present invention comprise dyes of Formulas 1to 6 which are hydrophilic and absorb light in the visible and nearinfrared regions of the electromagnetic spectrum. The blood clearancerate can be modified by formulating the dyes in liposomes, micelles, orother microparticles. This enhances their use for physiologicalmonitoring of many organs. The ease of modifying the clearance pathwaysof the dyes after in vivo administration permits their use forphysiological monitoring. Compounds with longer blood persistence areuseful for angiography and organ perfusion analysis, which isparticularly useful in organ transplant and critical ill patients.Predominant kidney clearance of the dyes enables their use for dynamicrenal function monitoring, and rapid liver uptake of the dyes from bloodserves as a useful index for the evaluation of hepatic function.

As illustrated in FIGS. 1-7, these dyes are designed to inhibitaggregation in solution by preventing intramolecular and intermolecularinduced hydrophobic interactions.

The present invention relates particularly to the novel compoundscomprising indoles of the general Formula 1

wherein R₃, R₄, R₅, R₆, and R₇, and Y₁ are independently selected fromthe group consisting of —H, C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl,C1-C20 polyhydroxyalkyl, C5-C20 polyhydroxyaryl, glucose derivatives ofR groups, saccharides, amino, C1-C10 aminoalkyl, cyano, nitro, halogen,hydrophilic peptides, arylpolysulfonates, C1-C10 alkyl, C1-C10 aryl,—SO₃T, —CO₂T, —OH, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T,(CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T,—(CH₂)_(a)CONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCO(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T,—(CH₂)_(a)OCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂,—(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂, —(CH₂)_(a)NHPO₃HT,—(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)CONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCONH(CH₂)_(b)PO₃HT, and—(CH₂)_(a)OCONH(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₁ is selected fromthe group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se; a, b,d, f, h, i, and j independently vary from 1-10; c, e, g, and kindependently vary from 1-100; R_(a), R_(b), R_(c), and R_(d) aredefined in the same manner as Y₁; T is either H or a negative charge.

The present invention also relates to the novel compounds comprisingbenzoindoles of general Formula 2

wherein R₈, R₉, R₁₀, R₁₁, R₁₂, R₁₃, R₁₄, and Y₂ are independentlyselected from the group consisting of —H, C1-C10 alkoxyl, C1-C10polyalkoxyalkyl, C1-C20 polyhydroxyalkyl, C5-C20 polyhydroxyaryl,glucose derivatives of R groups, saccharides, amino, C₁-C₁₀ aminoalkyl,cyano, nitro, halogen, hydrophilic peptides, arylpolysulfonates, C₁-C10alkyl, C1-C10 aryl, —SO₃T, —CO₂T, —OH, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T,—(CH₂)_(a)NHSO₃T, (CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T,—(CH₂)_(a)CONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCO(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T,—(CH₂)_(a)OCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂,—(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂, —(CH₂)_(a)NHPO₃HT,—(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)CONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCONH(CH₂)_(b)PO₃HT, and—(CH₂)_(a)OCONH(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₂ is selected fromthe group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se; a, b,d, f, h, i, and j independently vary from 1-10; c, e, g, and kindependently vary from 1-100; R_(a), R_(b), R_(c), and R_(d) aredefined in the same manner as Y₂; T is either H or a negative charge.

The present invention also relates to the novel compounds comprisingcyanine dyes of general Formula 3

wherein R₁₅, R₁₆, R₁₇, R₁₈, R₁₉, R₂₀, R₂₁, R₂₂, R₂₃, Y₃, and Z₃ areindependently selected from the group consisting of —H, C1-C10 alkoxyl,C1-C10 polyalkoxyalkyl, C1-C20 polyhydroxyalkyl, C5-C20 polyhydroxyaryl,glucose derivatives of R groups, saccharides, amino, C1-C10 aminoalkyl,cyano, nitro, halogen, hydrophilic peptides, arylpolysulfonates, C1-C10alkyl, C1-C10 aryl, —SO₃T, —CO₂T, —OH, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T,—(CH₂)_(a)NHSO₃T, (CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T,—(CH₂)_(a)CONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCO(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T,—CH₂)_(a)OCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂,—(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂, —(CH₂)_(a)NHPO₃HT,—(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)CONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCONH(CH₂)_(b)PO₃HT, and—(CH₂)_(a)OCONH(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₃ and X₃ are selectedfrom the group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se;V₃ is a single bond or is selected from the group consisting of —O—,—S—, —Se—, and —NR_(a); a, b, d, f, h, i, and j independently vary from1-10; c, e, g, and k independently vary from 1-100; a₃ and b₃ vary from0 to 5; R_(a), R_(b), R_(c), and R_(d) are defined in the same manner asY₃; T is either H or a negative charge.

The present invention further relates to the novel compounds comprisingcyanine dyes of general Formula 4

wherein R₂₄, R₂₅, R₂₆, R₂₇, R₂₈, R₂₉, R₃₀, R₃₁, R₃₂, R₃₃, R₃₄, R₃₅, R₃₆,Y₄, and Z₄ are independently selected from the group consisting of —H,C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl, C1-C20 polyhydroxyalkyl, C5-C20polyhydroxyaryl, glucose derivatives of R groups, saccharides, amino,C1-C10 aminoalkyl, cyano, nitro, halogen, hydrophilic peptides,arylpolysulfonates, C1-C10 alkyl, C1-C10 aryl, —SO₃T, —CO₂T, —OH,—(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T,(CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T,—(CH₂)_(a)CONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCO(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T,—CH₂)_(a)OCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂,—(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂, —(CH₂)_(a)NHPO₃HT,—(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)CONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCONH(CH₂)_(b)PO₃HT, and—(CH₂)_(a)OCONH(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₄ and X₄ are selectedfrom the group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se;V₄ is a single bond or is selected from the group consisting of —O—,—S—, —Se—, and —NR_(a); a₄ and b₄ vary from 0 to 5; a, b, d, f, h, i,and j independently vary from 1-10; c, e, g, and k independently varyfrom 1-100; R_(a), R_(b), R_(c), and R_(d) are defined in the samemanner as Y₄; T is either H or a negative charge.

The present invention also relates to the novel compounds comprisingcyanine dyes of general Formula 5

wherein R₃₇, R₃₈, R₃₉, R₄₀, R₄₁, R₄₂, R₄₃, R₄₄, R₄₅, Y₅, and Z₅ areindependently selected from the group consisting of —H, C1-C10 alkoxyl,C1-C10 polyalkoxyalkyl, C1-C20 polyhydroxyalkyl, C5-C20 polyhydroxyaryl,glucose derivatives of R groups, saccharides, amino, C1-C10 aminoalkyl,cyano, nitro, halogen, hydrophilic peptides, arylpolysulfonates, C1-C10alkyl, C1-C10 aryl, —SO₃T, —CO₂T, —OH, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T,—(CH₂)_(a)NHSO₃T, (CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T,—(CH₂)_(a)CONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCO(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T,—CH₂)_(a)OCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂,—(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂, —(CH₂)_(a)NHPO₃HT,—(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)CONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCONH(CH₂)_(b)PO₃HT, and—(CH₂)_(a)OCONH(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₅ and X₅ are selectedfrom the group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se;V₅ is a single bond or is selected from the group consisting of —O—,—S—, —Se—, and —NR_(a); D₅ is a single or a double bond; A₅, B₅ and E₅may be the same or different and are selected from the group consistingof —O—, —S—, —Se—, —P—, —NR_(a), —CR_(c)R_(d), CR_(c), alkyl, and —C═O;A₅, B₅, D₅, and E₅ may together form a 6 or 7 membered carbocyclic ringor a 6 or 7 membered heterocyclic ring optionally containing one or moreoxygen, nitrogen, or a sulfur atom; a, b, d, f, h, i, and jindependently vary from 1-10; c, e, g, and k independently vary from1-100; a₅ and b₅ vary from 0 to 5; R_(a), R_(b), R_(c), and R_(d) aredefined in the same manner as Y₅; T is either H or a negative charge.

The present invention also relates to the novel compounds comprisingcyanine dyes of general Formula 6

wherein R₄₆, R₄₇, R₄₈, R₄₉, R₅₀, R₅₁, R₅₂, R₅₃, R₅₄, R₅₅, R₅₆, R₅₇ andR₅₈, Y₆, and Z₆ are independently selected from the group consisting of—H, C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl, C1-C20 polyhydroxyalkyl,C5-C20 polyhydroxyaryl, glucose derivatives of R groups, saccharides,amino, C1-C10 aminoalkyl, cyano, nitro, halogen, hydrophilic peptides,arylpolysulfonates, C1-C10 alkyl, C1-C10 aryl, —SO₃T, —CO₂T, —OH,—(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T,(CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T,—(CH₂)_(a)CONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCO(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T,—CH₂)_(a)OCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂,—(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂, —(CH₂)_(a)NHPO₃HT,—(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)CONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCONH(CH₂)_(b)PO₃HT, and—(CH₂)_(a)OCONH(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₆ and X₆ are selectedfrom the group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se;V₆ is a single bond or is selected from the group consisting of —O—,—S—, —Se—, and —NR_(a); D₆ is a single or a double bond; A₆, B₆ and E₆may be the same or different and are selected from the group consistingof —O—, —S—, —Se—, —P—, —NR_(a), —CR_(c)R_(d), CR_(c), alkyl, and —C═O;A₆, B₆, D₆, and E₆ may together form a 6 or 7 membered carbocyclic ringor a 6 or 7 membered heterocyclic ring optionally containing one or moreoxygen, nitrogen, or sulfur atom; a, b, d, f, h, i, and j independentlyvary from 1-10; c, e, g, and k independently vary from 1-100; a₆ and b₆vary from 0 to 5; R_(a), R_(b), R_(c), and R_(d) are defined in the samemanner as Y₆; T is either H or a negative charge.

A hydrophilic chelate such as ethylenediaminetetraacetic acid (EDTA),diethylenetriaminepentaacetic acid (DPTA),1,4,7,10-tetraazacyclododecanetetraacetic acid (DOTA), or theirderivatives, can be attached to the compounds of Formulas 1-6 as one ormore R groups. These structures are expected to be highly water soluble.

The inventive compounds, compositions, and methods are advantageoussince they provide a real-time, accurate, repeatable measure of renalexcretion rate using exogenous markers under specific yet changingcircumstances. This represents a substantial improvement over anycurrently available or widely practiced method, since currently, noreliable, continuous, repeatable bedside method for the assessment ofspecific renal function by optical methods exists. Moreover, since theinventive method depends solely on the renal elimination of theexogenous chemical entity, the measurement is absolute and requires nosubjective interpretation based on age, muscle mass, blood pressure,etc. In fact it represents the nature of renal function in a particularpatient, under these particular circumstances, at a precise moment intime.

The inventive compounds, compositions, and methods provide simple,efficient, and effective monitoring of organ function. The compound isadministered and a sensor, either external or internal, is used todetect absorption and/or emission to determine the rate at which thecompound is cleared from the blood. By altering the R groups, thecompounds may be rendered more organ specific.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a reaction pathway for the preparation of indole derivatives.

FIG. 2 is a reaction pathway for the preparation of benzoindolederivatives.

FIG. 3 is a reaction pathway for the preparation of indocarbocyaninederivatives.

FIG. 4 is a reaction pathway for the preparation ofbenzoindocarbocyanine derivatives.

FIG. 5 is a reaction pathway for the preparation of robustindocarbocyanine derivatives.

FIG. 6 is a reaction pathway for the preparation of robustbenzoindocarbocyanine derivatives.

FIG. 7 is a reaction pathway for the preparation of long-wavelengthabsorbing indocarbocyanine derivatives.

FIG. 8 a is an absorption spectrum of indoledisulfonate in water.

FIG. 8 b is an emission spectrum of indoledisulfonate in water.

FIG. 9 a is an absorption spectrum of indocarbocyaninetetrasulfonate inwater.

FIG. 9 b is an emission spectrum of indocarbocyaninetetrasulfonate inwater.

FIG. 10 a is an absorption spectrum of chloroindocarbocyanine inacetonitrile.

FIG. 10 b is an emission spectrum of chloroindocarbocyanine inacetonitrile.

FIG. 11 is a blood clearance profile of carbocyanine-polyaspartic (10kDa) acid conjugate in a rat.

FIG. 12 is a blood clearance profile of carbocyanine-polyaspartic (30kDa) acid conjugate in a rat.

FIG. 13 is a blood clearance profile of indoledisulfonate in a rat.

FIG. 14 is a blood clearance profile of carbocyaninetetrasulfonates in arat.

DETAILED DESCRIPTION

In one embodiment the dyes of the invention serve as probes forcontinuous monitoring of renal function, especially for critically illpatients and kidney transplant patients.

In another embodiment, the dyes of the invention are useful for dynamichepatic function monitoring, especially for critically ill patients andliver transplant patients.

In another embodiment, the dyes of the invention are useful forreal-time determination of cardiac function, especially in patients withcardiac diseases.

In another embodiment, the dyes of the invention are useful formonitoring organ perfusion, especially for critically ill, cancer, andorgan transplant patients.

In another embodiment, the dyes are useful for assessing the functionalstatus of tumors and for monitoring tumor perfusion, such as in renal orhepatic cancer patients.

The novel dyes of the present invention are prepared according to themethods well known in the art, as illustrated in general in FIGS. 1-7and described for specific compounds in Examples 1-11.

In one embodiment, the novel compounds, also called tracers, of thepresent invention have the Formula 1, wherein R₃, R₄, R₅, R₆ and R₇, andY₁ are independently selected from the group consisting of —H, C1-C5alkoxyl, C1-C5 polyalkoxyalkyl, C1-C10 polyhydroxyalkyl, C5-C20polyhydroxyaryl, mono- and disaccharides, nitro, hydrophilic peptides,arylpolysulfonates, C1-C5 alkyl, C1-C10 aryl, —SO₃T, —CO₂T, —OH,—(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T,—(CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T,—CH₂(CH₂—O—CH₂)_(c)—CH₂—OH, —(CH₂)_(d)—CO₂T,—CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₁ is selected fromthe group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se; a, b,d, f, h, I, and j independently vary from 1-5; c, e, g, and kindependently vary from 1-20; R_(a), R_(b), R_(c), and R_(d) are definedin the same manner as Y₁; T is a negative charge.

In another embodiment, the novel compounds of the present invention havethe general Formula 2, wherein R₈, R₉, R₁₀, R₁₁, R₁₂, R₁₃, R₁₄, and Y₂are independently selected from the group consisting of —H, C1-C5alkoxyl, C1-C5 polyalkoxyalkyl, C1-C10 polyhydroxyalkyl, C5-C20polyhydroxyaryl, mono- and disaccharides, nitro, hydrophilic peptides,arylpolysulfonates, C1-C5 alkyl, C1-C10 aryl, —SO₃T, —CO₂T, —OH,—(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T,—(CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T,—CH₂(CH₂—O—CH₂)_(c)—CH₂—OH, —(CH₂)_(d)—CO₂T,—CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₂ is selected fromthe group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se; a, b,d, f, h, I, and j independently vary from 1-5; c, e, g, and kindependently vary from 1-20; R_(a), R_(b), R_(c), and R_(d) are definedin the same manner as Y₂; T is a negative charge.

In another embodiment, the novel compositions of the present inventionhave the general Formula 3, wherein R₁₅, R₁₆, R₁₇, R₁₈, R₁₉, R₂₀, R₂₁,R₂₂, R₂₃, Y₃, and Z₃ are independently selected from the groupconsisting of —H, C1-C5 alkoxyl, C1-C5 polyalkoxyalkyl, C1-C10polyhydroxyalkyl, C5-C20 polyhydroxyaryl, mono- and disaccharides,nitro, hydrophilic peptides, arylpolysulfonates, C1-C5 alkyl, C1-C10aryl, —SO₃T, —CO₂T, —OH, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T,—(CH₂)_(a)NHSO₃T, —(CH₂)_(a)CO₂(CH₂)_(b)SO₃T,—(CH₂)_(a)OCO(CH₂)_(b)SO₃T, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH, —(CH₂)_(d)—CO₂T,—CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₃ and X₃ are selectedfrom the group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se;V₃ is a single bond or is selected from the group consisting of —O—,—S—, —Se—, and —NR_(a); a, b, d, f, h, i, and j independently vary from1-5; c, e, g, and k independently vary from 1-50; a₃ and b₃ vary from 0to 5; R_(a), R_(b), R_(c), and R_(d) are defined in the same manner asY₃; T is either H or a negative charge.

In another embodiment, the novel compounds of the present invention havethe general Formula 4, wherein R₂₄, R₂₅, R₂₆, R₂₇, R₂₈, R₂₉, R₃₀, R₃₁,R₃₂, R₃₃, R₃₄, R₃₅, R₃₆, Y₄, and Z₄ are independently selected from thegroup consisting of —H, C1-C5 alkoxyl, C1-C5 polyalkoxyalkyl, C1-C10polyhydroxyalkyl, C5-C20 polyhydroxyaryl, mono- and disaccharides,nitro, hydrophilic peptides, arylpolysulfonates, C1-C5 alkyl, C1-C10aryl, —SO₃T, —CO₂T, —OH, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T,—(CH₂)_(a)NHSO₃T, —(CH₂)_(a)CO₂(CH₂)_(b)SO₃T,—(CH₂)_(a)OCO(CH₂)_(b)SO₃T, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH, —(CH₂)_(d)—CO₂T,—CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₄ and X₄ are selectedfrom the group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se;V₄ is a single bond or is selected from the group consisting of —O—,—S—, —Se—, and —NR_(a); a₄ and b₄ vary from 0 to 5; a, b, d, f, h, i,and j independently vary from 1-5; c, e, g, and k independently varyfrom 1-50; R_(a), R_(b), R_(c), and R_(d) are defined in the same manneras Y₄; T is either H or a negative charge.

In another embodiment, the novel compounds of the present invention havethe general Formula 5, wherein R₃₇, R₃₈, R₃₉, R₄₀, R₄₁, R₄₂, R₄₃, R₄₄,R₄₅, Y₅, and Z₅ are independently selected from the group consisting of—H, C1-C5 alkoxyl, C1-C5 polyalkoxyalkyl, C1-C10 polyhydroxyalkyl,C5-C20 polyhydroxyaryl, mono- and disaccharides, nitro, hydrophilicpeptides, arylpolysulfonates, C1-C5 alkyl, C1-C₁₀ aryl, —SO₃T, —CO₂T,—OH, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T,—(CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T,—CH₂(CH₂—O—CH₂)_(c)—CH₂—OH, —(CH₂)_(d)—CO₂T,—CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₅ and X₅ are selectedfrom the group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se;V₅ is a single bond or is selected from the group consisting of —O—,—S—, —Se—, and —NR_(a) D₅ is a single or a double bond; A₅, B₅ and E₅may be the same or different and are selected from the group consistingof —O—, —S—, —NR_(a), —CR_(c)R_(d), CR_(c), and alkyl; A₅, B₅, D₅, andE₅ may together form a 6 or 7 membered carbocyclic ring or a 6 or 7membered heterocyclic ring optionally containing one or more oxygen,nitrogen, or sulfur atom; a, b, d, f, h, i, and j independently varyfrom 1-5; c, e, g, and k independently vary from 1-50; a₅ and b₅ varyfrom 0 to 5; R_(a), R_(b), R_(c), and R_(d) are defined in the samemanner as Y₅; T is either H or a negative charge.

In another embodiment, the novel compounds of the present invention havethe general Formula 6, wherein R₄₆, R₄₇, R₄₈, R₄₉, R₅₀, R₅₁, R₅₂, R₅₃,R₅₄, R₅₅, R₅₆, R₅₇, R₅₈, Y₆, and Z₆ are independently selected from thegroup consisting of —H, C1-C5 alkoxyl, C1-C5 polyalkoxyalkyl, C1-C₁₀polyhydroxyalkyl, C5-C20 polyhydroxyaryl, mono- and disaccharides,nitro, hydrophilic peptides, arylpolysulfonates, C1-C5 alkyl, C1-C₁₀aryl, —SO₃T, —CO₂T, —OH, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T,—(CH₂)_(a)NHSO₃T, —(CH₂)_(a)CO₂(CH₂)_(b)SO₃T,—(CH₂)_(a)OCO(CH₂)_(b)SO₃T, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH, —(CH₂)_(d)—CO₂T,—CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₆ and X₆ are selectedfrom the group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se;V₆ is a single bond or is selected from the group consisting of —O—,—S—, —Se—, and —NR_(a); D₆ is a single or a double bond; A₆, B₆ and E₆may be the same or different and are selected from the group consistingof —O—, —S—, —NR_(a), —CR_(c)R_(d), CR_(c), and alkyl; A₆, B₆, D₆, andE₆ may together form a 6 or 7 membered carbocyclic ring or a 6 or 7membered heterocyclic ring optionally containing one or more oxygen,nitrogen, or sulfur atom; a, b, d, f, h, i, and j independently varyfrom 1-5; c, e, g, and k independently vary from 1-50; a₅ and b₅ varyfrom 0 to 5; R_(a), R_(b), R_(c), and R_(d) are defined in the samemanner as Y₆; T is either H or a negative charge.

The compounds of the invention can be formulated into diagnostic andtherapeutic compositions for enteral or parenteral administration. Thesecompositions contain an effective amount of the dye along withconventional pharmaceutical carriers and excipients appropriate for thetype of administration contemplated. For example, parenteralformulations advantageously contain the inventive agent in a sterileaqueous solution or suspension. Parenteral compositions may be injecteddirectly or mixed with a large volume parenteral composition forsystemic administration. Such solutions also may containpharmaceutically acceptable buffers and, optionally, electrolytes suchas sodium chloride.

Formulations for enteral administration may vary widely, as is wellknown in the art. In general, such formulations are liquids, whichinclude an effective amount of the inventive agent in aqueous solutionor suspension. Such enteral compositions may optionally include buffers,surfactants, thixotropic agents, and the like. Compositions for oraladministration may also contain flavoring agents and other ingredientsfor enhancing their organoleptic qualities.

The compositions are administered in doses effective to achieve thedesired effect or result. The dosage of the tracers may vary accordingto the clinical procedure contemplated and generally ranges from 1picomolar to 100 millimolar. The compositions may be administered to apatient, typically a warm-blooded animal either systemically or locallyto the organ or tissue to be imaged, and the patient then subject to theimaging procedure. The tracers may be administered to the patient by anysuitable method, including intravenous, intraperitoneal, or subcutaneousinjection or infusion, oral administration, transdermal absorptionthrough the skin, aerosols, or by inhalation. The detection of thetracers is achieved by optical fluorescence, absorbance, or lightscattering methods known in the art (Muller et al. Eds., Medical OpticalTomography, SPIE Volume IS11, 1993, which is expressly incorporatedherein by reference) using invasive or non-invasive probes such asendoscopes, catheters, ear clips, hand bands, surface coils, fingerprobes, and the like. Physiological function is correlated with theclearance profiles and rates of these agents from body fluids (Dorshowet al., Non-Invasive Fluorescence Detection of Hepatic and RenalFunction, Bull. Am. Phys. Soc. 1997, 42, 681, which is expresslyincorporated by reference herein).

The inventive composition may be administered for imaging by more thanone modality. As one example, the composition may be used for imaging byoptical imaging alone, by nuclear imaging alone, or by both optical andnuclear imaging modalities when a radioactive isotope is included in thechemical formula, such as replacing a halogen atom with a radioactivehalogen, and/or including a radioactive metal ion such as Tc⁹⁹, In¹¹¹,etc. As another example, the composition may be used for imaging byoptical imaging alone, by magnetic resonance (MR) alone, or by bothoptical and MR modalities when a paramagnetic metal ion such asgadolinium or manganese is included in the chemical formula.

It will also be appreciated that the inventive compositions may beadministered with other contrast agents or media used to enhance animage from a non-optical modality. These include agents for enhancing animage obtained by modalities including but not limited to MR, ultrasound(US), x-ray, positron emission tomography (PET), computed tomography(CT), single photon emission computed tomography (SPECT), optoacoustic(e.g. U.S. Pat. Nos. 5,840,023 and 5,977,538 which are expresslyincorporated by reference herein in their entirety), etc. Both opticaland non-optical agents may be formulated as a single composition (thatis, one composition containing one, two, or more components, forexample, an optical agent and a MR agent), or may be formulated asseparate compositions. The inventive optical imaging contrast agent andthe non-optical contrast agent are administered in doses effective toachieve the desired enhancement, diagnosis, therapy, etc., as known toone skilled in the art. The inventive compositions, either alone orcombined with a contrast agent, may be administered to a patient,typically a warm-blooded animal, systemically or locally to the organ ortissue to be imaged. The patient is then imaged by optical imagingand/or by another modality. As one example of this embodiment, theinventive compounds may be added to contrast media compositions. Asanother example, the inventive compositions may be co-administered withcontrast media, either simultaneously or within the same diagnosticand/or therapeutic procedure (for example, administering the inventivecomposition and administering a contrast agent then performing opticalimaging followed by another imaging modality, or administering theinventive composition and administering a contrast agent then performinganother imaging modality followed by optical imaging, or administeringthe inventive composition and optical imaging, then administering acontrast agent and MR, US, CT, etc. imaging, or administering a contrastagent and imaging by MR, US, CT, etc., then administering the inventivecomposition and optical imaging, or administering the inventivecomposition and a contrast agent, and simultaneously imaging by anoptical modality and MR, US, CT, etc.). As another example, an opticalimaging agent may be added as an additive or excipient for a non-opticalimaging modality. In this embodiment, the optically active component,such as the dyes disclosed herein, could be added as a buffering agentto control pH or as a chelate to improve formulation stability, etc. inMR contrast media, CT contrast media, x-ray contrast media, US contrastmedia, etc. The MR, CT, x-ray, US contrast media would then alsofunction as an optical imaging agent. The information obtained from themodality using the non-optical contrast agent is useful in combinationwith the image obtained using the optical contrast agent.

In one embodiment, the agents may be formulated as micelles, liposomes,microcapsules, or other microparticles. These formulations may enhancedelivery, and localization of the inventive compounds to/at the desiredorgan or site. The target specificity of these formulations can beenhanced by using suitable targeting molecules such as peptides,saccharides, fatty acids, etc. Preparation and loading of these are wellknown in the art.

As one example, liposomes may be prepared from dipalmitoylphosphatidylcholine (DPPC) or egg phosphatidylcholine (PC) because thislipid has a low heat transition. Liposomes are made using standardprocedures as known to one skilled in the art (e.g., Braun-Falco et al.,(Eds.), Griesbach Conference, Liposome Dermatics, Springer-Verlag,Berlin (1992)). Polycaprolactone, poly(glycolic) acid, poly(lactic)acid, polyanhydride or lipids may be formulated as microspheres. As anillustrative example, the optical agent may be mixed with polyvinylalcohol (PVA), the mixture then dried and coated with ethylene vinylacetate, then cooled again with PVA. In a liposome, the optical agentmay be within one or both lipid bilayers, in the aqueous between thebilayers, or with the center or core. Liposomes may be modified withother molecules and lipids to form a cationic liposome. Liposomes mayalso be modified with lipids to render their surface more hydrophilicwhich increases their circulation time in the bloodstream. Thethus-modified liposome has been termed a “stealth” liposome, or along-lived liposome, as described in U.S. Pat. Nos. 6,277403; 6,610,322;5,631,018; 5,395,619; and 6,258,378, each of which is expresslyincorporated by reference herein in its entirety, and in StealthLiposomes, Lasic and Martin (Eds.) 1995, CRC Press, London.Encapsulation methods include detergent dialysis, freeze drying, filmforming, injection, as known to one skilled in the art and disclosed in,for example, U.S. Pat. No. 6,406,713 which is expressly incorporated byreference herein in its entirety.

The agent formulated in liposomes, microcapsules, etc. may beadministered by any of the routes previously described. In a formulationapplied topically, the optical agent is slowly released over time. In aninjectable formulation, the liposome capsule circulates in thebloodstream and is delivered to a desired site.

As another example, microparticles such as ultra small iron oxideparticles (USPIO) and other metallic particles such as silver or goldparticles coated with or attached (covalently or non-covalently) withthe inventive compounds may be used for optical imaging and/or MRI. Suchparticles are known to one skilled in the art as disclosed in, forexample, U.S. Pat. No. 5,492,814 and Journal of Biomedical Optics 8(3),472-478 (July 2003) which are expressly incorporated by reference hereinin their entirety.

Organ function can be assessed either by the differences in the mannerin which the normal and impaired cells remove the tracer from thebloodstream, by measuring the rate or accumulation of these tracers inthe organs or tissues, or by obtaining tomographic images of the organsor tissues. Blood pool clearance may be measured non-invasively fromconvenient surface capillaries such as those found in an ear lobe or afinger, for example, using an ear clip or finger clip sensor, or may bemeasured invasively using an endovascular catheter. Accumulation of thetracer within the cells of interest can be assessed in a similarfashion. The clearance of the tracer dyes may be determined by selectingexcitation wavelengths and filters for the emitted photons. Theconcentration-time curves may be analyzed in real time by amicroprocessor. In order to demonstrate feasibility of the inventivecompounds to monitor organ function, a non-invasive absorbance orfluorescence detection system to monitor the signal emanating from thevasculature infused with the compounds is used. Indole derivatives, suchas those of Formulas 1-6, fluoresce at a wavelength between 350 nm and1300 nm when excited at the appropriate wavelength as is known to, orreadily determined by, one skilled in the art.

In addition to the noninvasive techniques, a modified pulmonary arterycatheter can be used to make the necessary measurements (Dorshow et al.,Monitoring Physiological Function by Detection of Exogenous FluorescentContrast Agents, in Optical Diagnostics of Biological Fluids IV,Priezzhev and Asakura, Editors, Procedings of SPIE 1999, 3599, 2-8,which is expressly incorporated by reference herein). Currently,pulmonary artery catheters measure only intravascular pressures, cardiacoutput and other derived measures of blood flow. Critically ill patientsare managed using these parameters, but rely on intermittent bloodsampling and testing for assessment of renal function. These laboratoryparameters represent discontinuous data and are frequently misleading inmany patient populations. Yet, importantly, they are relied upon heavilyfor patient assessment, treatment decisions, and drug dosing.

The modified pulmonary artery catheter incorporates an optical sensorinto the tip of a standard pulmonary artery catheter. This wavelengthspecific optical sensor can monitor the renal function specificelimination of an optically detectable chemical entity. Thus, by amethod analogous to a dye dilution curve, real-time renal function canbe monitored by the disappearance of the optically detected compound.Modification of a standard pulmonary artery catheter only requiresmaking the fiber optic sensor wavelength specific, as is known to oneskilled in this art. Catheters that incorporate fiber optic technologyfor measuring mixed venous oxygen saturation currently exist.

The present invention may be used for rapid bedside evaluation of renalfunction and also to monitor the efficiency of hemodialysis. Theinvention is further demonstrated by the following examples. Since manymodifications, variations, and changes in detail may be made to thedescribed embodiments, it is intended that all matter in the foregoingdescription and shown in the accompanying drawings be interpreted asillustrative and not limiting.

EXAMPLE 1 Synthesis of Indole Disulfonate FIG. 1, Compound 5, Y₇═SO₃ ⁻;X₇═H; n=1

A mixture of 3-methyl-2-butanone (25.2 mL), andp-hydrazinobenzenesulfonic acid (15 g) in acetic acid (45 mL) was heatedat 110° C. for 3 hours. After reaction, the mixture was allowed to coolto room temperature and ethyl acetate (100 mL) was added to precipitatethe product, which was filtered and washed with ethyl acetate (100 mL).The intermediate compound, 2,3,3-trimethylindolenium-5-sulfonate (FIG.1, compound 3) was obtained as a pink powder in 80% yield. A portion ofcompound 3 (9.2 g) in methanol (115 mL) was carefully added to asolution of KOH in isopropanol (100 mL). A yellow potassium salt of thesulfonate was obtained in 85% yield after vacuum-drying for 12 hours. Aportion of the 2,3,3-trimethylindolenium-5-sulfonate potassium salt (4g) and 1,3-propanesultone (2.1 g) was heated in dichlorobenzene (40 mL)at 110° C. for 12 hours. The mixture was allowed to cool to roomtemperature and the resulting precipitate was filtered and washed withisopropanol. The resulting pink powder was dried under vacuum to give97% of the desired compound.

Other compounds prepared by a similar method described above includepolyhydroxyl indoles such as

EXAMPLE 2 Synthesis of Indole Disulfonate FIG. 1, Compound 5, Y₇═SO₃ ⁻,X₇═H; n=2

This compound was prepared by the same procedure described in Example 1,except that 1,4-butanesultone was used in place of 1,3-propanesultone.

EXAMPLE 3 Synthesis of Benzoindole Disulfonate FIG. 2, Compound 8,Y₇,Y₈═SO₃ ⁻, X₇═H; n=2

This compound was prepared by the same procedure described in Example 1,except that hydrazinonaphthalenedisulfonic acid was used in place ofhydrazinobenzenesulfonic acid.

Other compounds prepared by a similar method include polyhydroxyindolessuch as:

EXAMPLE 4 Synthesis of Benzoindole Disulfonate FIG. 2, Compound 8,Y₇,Y₈═SO₃ ⁻, X₇═OH; n=4

This compound was prepared by the same procedure described in Example 1,except that 3-hydroxymethyl-4-hydroxyl-2-butanone was used in place of3-methyl-2-butanone.

EXAMPLE 5 Synthesis of Bis(ethylcarboxymethyl)indocyanine Dye

A mixture of 1,1,2-trimethyl-[1H]-benz[e]indole (9.1 g, 43.58 mmoles)and 3-bromopropanoic acid (10.0 g, 65.37 mmoles) in 1,2-dichlorobenzene(40 mL) was heated at 110° C. for 12 hours. The solution was cooled toroom temperature and the red residue obtained was filtered and washedwith acetonitrile: diethyl ether (1:1) mixture. The solid obtained wasdried under vacuum to give 10 g (64%) of light brown powder. A portionof this solid (6.0 g; 16.56 mmoles), glutaconaldehyde dianilmonohydrochloride (2.36 g, 8.28 mmoles) and sodium acetate trihydrate(2.93 g, 21.53 mmoles) in ethanol (150 mL) were refluxed for 90 minutes.After evaporating the solvent, 40 mL of 2 N aqueous HCl was added to theresidue and the mixture was centrifuged and the supernatant wasdecanted. This procedure was repeated until the supernatant becamenearly colorless. About 5 mL of water: acetonitrile (3:2) mixture wasadded to the solid residue and lyophilized to obtain 2 g of dark greenflakes. The purity of the compound was established with ¹H-NMR andliquid chromatography/mass spectrometry (LC/MS).

EXAMPLE 6 Synthesis of Bis(pentylcarboxymethyl)indocyanine Dye

A mixture of 2,2,3-trimethyl-[1H]-benz[e]indole (20 g, 95.6 mmoles) and6-bromohexanoic acid (28.1 g, 144.1 mmoles) in 1,2-dichlorobenzene (250mL) was heated at 110 C for 12 hours. The green solution was cooled toroom temperature and the brown solid precipitate formed was collected byfiltration. After washing the solid with 1,2-dichlorobenzene and diethylether, the brown powder obtained (24 g, 64%) was dried under vacuum atroom temperature. A portion of this solid (4.0 g; 9.8 mmoles),glutaconaldehyde dianil monohydrochloride (1.4 g, 5 mmoles) and sodiumacetate trihydrate (1.8 g, 12.9 mmoles) in ethanol (80 mL) were refluxedfor 1 hour. After evaporating the solvent, 20 mL of a 2 N aqueous HClwas added to the residue and the mixture was centrifuged and thesupernatant was decanted. This procedure was repeated until thesupernatant became nearly colorless. About 5 mL of water:acetonitrile(3:2) mixture was added to the solid residue and lyophilized to obtainabout 2 g of dark green flakes. The purity of the compound wasestablished with ¹H-NMR, HPLC, and LC-MS.

EXAMPLE 7 Synthesis of Polyhydroxyindole Sulfonate FIG. 3, Compound 13,Y₇,Y₈═O₃ ⁻, X₇═OH; n=2

Phosphorus oxychloride (37 ml, 0.4 mole) was added dropwise withstirring to a cooled (−2° C.) mixture of dimethylformamide (DMF, 0.5mole, 40 mL) and dichloromethane (DCM, 40 mL), followed by the additionof acetone (5.8 g, 0.1 mole). The ice bath was removed and the solutionrefluxed for 3 hours. After cooling to room temperature, the product waseither partitioned in water/DCM, separated and dried, or was purified byfractional distillation. Nuclear magnetic resonance and mass spectralanalyses showed that the desired intermediate, 10, was obtained.Reaction of the intermediate with 2 equivalents of2,2,3-trimethyl-[H]-benz[e]indolesulfonate-N-propanoic acid and 2equivalents of sodium acetate trihydrate in ethanol gave a blue-greensolution after 1.5 hours at reflux. Further functionalization of the dyewith bis(isopropylidene)acetal protected monosaccharide is effected bythe method described in the literature (Flanagan et al., Near infraredheavy-atom-modified fluorescent dyes for base-calling in DNA-sequencingapplication using temporal discrimination. Anal. Chem., 1998, 70(13),2676-2684).

EXAMPLE 8 Synthesis of Polyhydroxyindole Sulfonate FIG. 4, Compound 16,Y₇,Y₈═SO₃ ⁻, X₇═H; n=1

Preparation of this compound was readily accomplished by the sameprocedure described in Example 6 using p-hydroxybenzenesulfonic acid inthe place of the monosaccharide, and benzoindole instead of indolederivatives.

EXAMPLE 9 Synthesis of Polyhydroxyindole Sulfonate FIG. 5, Compound 20,Y₇,Y₈═H; X₇═OH; n=1

The hydroxyindole compound was readily prepared by a literature method(Southwick et a., One pot Fischer synthesis of(2,3,3-trimethyl-3-H-indol-5-yl)-acetic acid derivatives asintermediates for fluorescent biolabels, Org. Prep. Proced. Int. Briefs,1988, 20(3), 279-284). Reaction of p-carboxymethylphenylhydrazinehydrochloride (30 mmol, 1 equiv.) and 1,1-bis(hydroxymethyl)propanone(45 mmol, 1.5 equiv.) in acetic acid (50 mL) at room temperature for 30minutes and at reflux for 1 gave(3,3-dihydroxymethyl2-methyl-3-H-indol-5-yl)-acetic acid as a solidresidue.

The intermediate 2-chloro-1-formyl-3-hydroxymethylenecyclo-hexane wasprepared as described in the literature (Reynolds and Drexhage, Stableheptamethine pyrylium dyes that absorb in the infrared. J. Org. Chem.,1977, 42(5), 885-888). Equal volumes (40 mL each) of dimethylformamide(DMF) and dichloromethane were mixed and the solution was cooled to −10°C. in acetone-dry ice bath. Under argon atmosphere, phosphorusoxychloride (40 mL) in dichloromethane was added dropwise to the coolDMF solution, followed by the addition of 10 g of cyclohexanone. Theresulting solution was allowed to warm up to room temperature and heatedat reflux for 6 hours. After cooling to room temperature, the mixturewas poured into ice-cold water and stored at 4° C. for 12 hours. Ayellow powder was obtained. Condensation of a portion of this cyclicdialdehyde (1 equivalent) with the indole intermediate (2 equivalents)was carried out as described in Example 5. Further, thefunctionalization of the dye with bis (isopropylidene)acetal protectedmonosaccharide was effected by the method described in the literature(Flanagan et al., Near infrared heavy-atom-modified fluorescent dyes forbase-calling in DNA-sequencing application using temporaldiscrimination. Anal. Chem., 1998, 70(13), 2676-2684).

EXAMPLE 10 Synthesis of Polyhydroxylbenzoindole Sulfonate FIG. 6,Compound 22, Y₇, Y₈═H; X₇═OH; n=1

A similar method described in Example 8 was used to prepare thiscompound by replacing the indole with benzoindole derivatives.

EXAMPLE 11 Synthesis of Rigid Heteroatomic Indole Sulfonate FIG. 7,Compound 27, Y₇,Y₈,X₇═H; n=1

Starting with 3-oxo-4-cyclohexenone, this heteroatomic hydrophilic dyewas readily prepared as described in Example 8.

EXAMPLE 12 Minimally Invasive Monitoring of the Blood Clearance Profileof the Dyes

A laser of appropriate wavelength for excitation of the dye chromophorewas directed into one end of a fiber optic bundle and the other end waspositioned a few millimeters from the ear of a rat. A second fiber opticbundle was also positioned near the same ear to detect the emittedfluorescent light, and the other end was directed into the optics andelectronics for data collection. An interference filter (IF) in thecollection optics train was used to select emitted fluorescent light ofthe appropriate wavelength for the dye chromophore.

Sprague-Dawley or Fischer 344 rats were anesthetized with urethaneadministered via intraperitoneal injection at a dose of 1.35 g/kg bodyweight. After the animals had achieved the desired plane of anesthesia,a 21 gauge butterfly with 12″ tubing was placed in the lateral tail veinof each animal and flushed with heparinized saline. The animals wereplaced onto a heating pad and kept warm throughout the entire study. Thelobe of the left ear was affixed to a glass microscope slide to reducemovement and vibration.

Incident laser light delivered from the fiber optic was centered on theaffixed ear. Data acquisition was then initiated, and a backgroundreading of fluorescence was obtained prior to administration of the testagent.

The compound was administered to the animal through a bolus injection inthe lateral tail vein. The dose was typically 0.05 to 20 μmole/kg ofbody weight. The fluorescence signal rapidly increased to a peak value,then decayed as a function of time as the conjugate cleared from thebloodstream.

This procedure was repeated with several dye-peptide conjugates innormal and tumored rats. Representative profiles are shown in FIGS.6-10.

While the invention has been disclosed by reference to the details ofpreferred embodiments of the invention, it is to be understood that thedisclosure is intended in an illustrative rather than in a limitingsense, as it is contemplated that modifications will readily occur tothose skilled in the art, within the spirit of the invention and thescope of the appended claims.

1. A compound of Formula 2 for use in a diagnostic or therapeuticprocedure,

wherein W₂ is —CR_(c)R_(d); Y₂ is selected from the group consisting ofC1-C10 alkoxyl, C1-C10 polyalkoxyalkyl, C1-C20 polyhydroxyalkyl, C5-C20polyhydroxyaryl, saccharides, C1-C10 aminoalkyl, hydrophilic peptides,arylpolysulfonates, C1-C10 aryl, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T,—(CH₂)_(a)NHSO₃T, —(CH₂)_(a)CO₂(CH₂)_(b)SO₃T,—(CH₂)_(a)OCO(CH₂)_(b)SO₃T, —(CH₂)_(a)CONH(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCO(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T, —(CH₂)_(a)OCONH(CH₂)_(b)SO₃T,—(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂, —(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂,—(CH₂)_(a)NHPO₃HT, —(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)CONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCONH(CH₂)_(b)PO₃HT, and—(CH₂)_(a)OCONH(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(I)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—(CH₂—CO₂T; R₈ is C1-C10 alkyl;R₉, R₁₀, R₁₁, R₁₂, R₁₃ and R₁₄ are independently selected from the groupconsisting of H, C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl, C1-C20polyhydroxyalkyl, C5-C20 polyhydroxyaryl, saccharides, amino, C1-C10aminoalkyl, cyano, nitro, halogen, hydrophilic peptides,arylpolysulfonates, C1-C10 alkyl, C1-C10 aryl; —SO₃T, —CO₂T, —OH,—(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T,—(CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T,—(CH₂)_(a)CONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCO(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T,—(CH₂)_(a)OCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂,—(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂, —(CH₂)_(a)NHPO₃HT,—(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)CONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCONH(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(I)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; a, b, d, f, h, i, andj independently vary from 1-10; c, e, g, and k independently vary from1-100; R_(a), R_(b), R_(c), and R_(d) are defined in the same manner asY₂; and T is either H or a negative charge.
 2. The compound of claim 1wherein R₈ is C1 alkyl.
 3. The compound of claim 1 wherein R₉ to R₁₄ is—H or —SO₃T.
 4. The compound of claim 1 wherein Y₂ is selected from thegroup consisting of —(CH₂)_(d)—CO₂T, —(CH₂)_(a)NHSO₃T, —(CH₂)_(a)SO₃T,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, and —(CH₂)_(h)—N(R_(a))—(CH₂)_(I)—CO₂T. 5.The compound of claim 1 wherein the compound comprises a radioactivehalogen.
 6. A method for performing a diagnostic or therapeuticprocedure comprising administering to a mammal an effective amount of acompound of formula 2

wherein W₂ is —CRcRd; Y₂ is independently selected from the groupconsisting of C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl, C1-C20polyhydroxyalkyl, C5-C20 polyhydroxyaryl, saccharides, C1-C10aminoalkyl, hydrophilic peptides, arylpolysulfonates, C1-C10 aryl,—(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T,—(CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T,—(CH₂)_(a)CONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCO(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T,—(CH₂)_(a)OCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂,—(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂, —(CH₂)_(a)NHPO₃HT,—(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)CONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCONH(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(I)—CO₂T, and—(CH₂)_(j)—N(R_(b)) —CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; R₈ is C1-C10 alkyl;R₉, R₁₀, R₁₁, R₁₂, R₁₃ and R₁₄ are independently selected from the groupconsisting of H, C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl, C1-C20polyhydroxyalkyl, C5-C20 polyhydroxyaryl, saccharides, amino, C1-C10aminoalkyl, cyano, nitro, halogen, hydrophilic peptides,arylpolysulfonates, C1-C10 alkyl, C1-C10 aryl; —SO₃T, —CO₂T, —OH,—(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T,—(CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T,—(CH₂)_(a)CONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCO(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T,—(CH₂)_(a)OCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂,—(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂, —(CH₂)_(a)NHPO₃HT,—(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)CONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCONH(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(I)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; a, b, d, f, h, i, andj independently vary from 1-10; c, e, g, and k independently vary from1-100; R_(a), R_(b), R_(c), and R_(d) are defined in the same manner asY₂; and T is either H or a negative charge; and thereafter performingthe diagnostic or therapeutic procedure.
 7. The method of claim 6wherein the compound of formula 2 R₈ is C1 alkyl.
 8. The method of claim6 wherein compound of formula 2 R₉ to R₁₄ is —H or —SO₃T.
 9. The methodof claim 6 wherein compound of formula 2 Y₂ is selected from—(CH₂)_(d)—CO₂T, —(CH₂)_(a)NHSO₃T, —(CH₂)_(a)SO₃T,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, and —(CH₂)_(h)—N(R_(a))—(CH₂)_(I)—CO₂T.10. The method of claim 6 wherein the compound of formula 2 comprises aradioactive halogen.
 11. The method of claim 6 wherein the compoundadministered further comprises a radioactive metal ion or a paramagneticmetal ion.
 12. The method of claim 6 further comprising imaging by atleast one of optical imaging, nuclear imaging, or magnetic resonanceimaging.
 13. The method of claim 6 wherein the compound is administeredin a formulation selected from at least one of liposomes, micelles,microcapsules, or microparticles.
 14. The method of claim 6 wherein thecompound is administered in a formulation selected from at least one ofultra small iron oxide particles, silver particles, or gold particles.15. The method of claim 6 further comprising administering a non-opticalcontrast agent and imaging by at least one of magnetic resonance,ultrasound, x-ray, position emission tomography, computed tomography,optoacoustic imaging, and single photon emission computed tomography.16. The method of claim 6 wherein the procedure is for physiologicalfunction monitoring selected from least one of renal functionmonitoring, cardiac function monitoring, or kidney function monitoring,thereby determining organ perfusion in vivo.
 17. The method of claim 6further comprising optically imaging the mammal.
 18. A method of imaginga patient in need thereof, the method comprising administering anon-optical contrast agent composition further comprising the compoundof any of claim 1, and performing at least one of an optical imagingprocedure or a non-optical imaging procedure.
 19. The method of claim 18wherein the non-optical contrast agent composition is selected from atleast one of a magnetic resonance composition, a computed tomographycomposition, an x-ray composition, a nuclear imaging composition, apositron emission tomography composition, a single photon emissioncomputed tomography composition, an optoacoustic composition, or anultrasound composition.
 20. The method of claim 18 wherein the compoundstabilizes or buffers the non-optical contrast agent.